title: Ezrin/radixin/moesin (ERM) proteins bind to a positively charged amino acid cluster in the juxta-membrane cytoplasmic domain of CD44, CD43, and ICAM-2.
abstract: CD44 has been identified as a membrane-binding partner for ezrin/radixin/moesin (ERM) proteins, plasma membrane/actin filament cross-linkers. ERM proteins, however, are not necessarily colocalized with CD44 in tissues, but with CD43 and ICAM-2 in some types of cells. We found that glutathione-S-transferase fusion proteins with the cytoplasmic domain of CD43 and ICAM-2, as well as CD44, bound to moesin in vitro. The regions responsible for the in vitro binding of CD43 and CD44 to moesin were narrowed down to their juxta-membrane 20-30-amino acid sequences in the cytoplasmic domain. These sequences and the cytoplasmic domain of ICAM-2 (28 amino acids) were all characterized by the positively charged amino acid clusters. When E-cadherin chimeric molecules bearing these positively charged amino acid clusters of CD44, CD43, or ICAM-2 were expressed in mouse L fibroblasts, they were co-concentrated with ERM proteins at microvilli, whereas those lacking these clusters were diffusely distributed on the cell surface. The specific binding of ERM proteins to the juxta-membrane positively charged amino acid clusters of CD44, CD43, and ICAM-2 was confirmed by immunoprecipitation and site-directed mutagenesis. From these findings, we conclude that ERM proteins bind to integral membrane proteins bearing a positively charged amino acid cluster in their juxta-membrane cytoplasmic domain.
authors.affiliation: Department of Cell Biology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan. yonemura@mfour.med.kyoto-u.ac.jp
authors.forename: S
authors.initials: S
authors.lastname: Yonemura
authors.forename: M
authors.initials: M
authors.lastname: Hirao
authors.forename: Y
authors.initials: Y
authors.lastname: Doi
authors.forename: N
authors.initials: N
authors.lastname: Takahashi
authors.forename: T
authors.initials: T
authors.lastname: Kondo
authors.forename: S
authors.initials: S
authors.lastname: Tsukita
authors.forename: S
authors.initials: S
authors.lastname: Tsukita
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D001798:Blood Proteins / Q000378:metabolism*; D015820:Cadherins / Q000032:analysis / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D003598:Cytoskeletal Proteins; D005982:Glutathione Transferase / Q000235:genetics; D018960:Hyaluronan Receptors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007739:L Cells; D051917:Leukosialin; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins; D008871:Microvilli / Q000737:chemistry; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012795:Sialoglycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000235:genetics / Q000502:physiology; D000099149:Ezrin
chemical_list: D000596:Amino Acids; D015703:Antigens, CD; D001798:Blood Proteins; D015820:Cadherins; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D005982:Glutathione Transferase; D018960:Hyaluronan Receptors; D051917:Leukosialin; D008565:Membrane Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D012795:Sialoglycoproteins; D000099149:Ezrin; C065289:ICAM2 protein, human; C497863:Spn protein, mouse; C071058:moesin; C071776:radixin